Objective To study the effects of Nitric oxide(NO),Nitric oxide synthases (NOS)on calcium-activated potassium channels (KCa) of mesenteric artery smooth muscle(SMC)between patients with essential hypertension (EH) and normotensive patients.Methods (1)Mesenteric artery branch from 21 EH and 18 normotensive patients respectively was digested by enzyme.Patch clamp technique was used to pull cell-attached and inside-out patches on mesenteric artery SMC from EH.The signal channel open number probability (Po),open dwell-time(To) and close dwell- time(Tc),were recorded.(2)The levels of plasma NO?NOS were measured by colorimetry analysis method.(3)The analysis of linear correlation was performed to estimate the relationship between the levels of plasma NO and Po?To?Tc of KCa of mesenteric artery SMC in the subjects of the two groups.Results (1)Compared to that of normotensive patients,the activities of KCa channels of patients with EH was higher.After adding Ca 2+ to cytoplasm,the Po of KCa channels in normotensive patients increased significantly.But there were few changes in EH group.(2)The levels of plasma NO?NOS were lower in EH group than in control group.(3)Linear correlation analysis showed that the levels of plasma NO were correlated positively with Po?To in normotensive patients,wherese the positive correlation were decreased in EH group.Conclusion The activities of KCa channels of patients with EH increase significantly.but the sensitivity to Ca 2+ decreased.NO?NOS may stimulate the KCa channels of normotensive and EH patients,but they may not be main factors in EH patients.

Objective To study the effects of high matrix metalloproteinase 9 (MMP 9 ) on the biological behaviors of fibroblasts,in order to clarify the possible mechanisms in the wound healing of diabetic foot.Methods Establish the cell model of skin fibroblast with high expression of MMP 9 by high glucose (22.0 mmol/L) and high homocysteine (100 μmol/L) co-culture.Control group was incubated with normal glucose (5.5 mmol/L).Realtime PCR,ELISA and gelatin zymography were used to detect the MMP 9 mRNA,protein expression and activity of MMP 9.Flow cytometry,CCK-8,ELISA assay,scratch test and transwell were used to detect cell proliferation,viability,collagen (hydroxyproline) secretion,horizontal migration and vertical migration of cells.Results Data were expressed as (x- ± s).Differences were considered significant if their P value was ＜ 0.05.Results The expression of MMP 9 mRNA,protein levels and the activity of MMP 9 in high MMP 9 group were 6.05 folds,4.12 folds and 1.58 folds higher than those in control group (P ＜ 0.01,respectively).The proportion of S phase cells,proliferation index,cell viability,collagen (hydroxyproline) secretion,6 h horizontal migration rate and the number of vertical migration cells in high MMP9 group decreased by 29.8 ％,18.1％,23.3 ％,68.7 ％,45.0 ％ and 21.4 ％ than those in control group (P ＜ 0.01,respectively ). Conclusions Fibroblast with high expression of MMP 9 have decreased proliferation,activity,collagen secretion and reduced migration,which suggests that MMP 9 may inhibit the biological behaviors of fibroblasts.

Objective To determine the involvement of DNA demethylation in tumor necrosis factor-α(TNF-α)-induced matrix metalloproteinase 9 ( MMP9) expression in human epidermal keratinocytes. Methods Real-time RT-PCR, Western blot, and enzyme-linked immuno sorbent assay (ELISA) were performed to determine the mRNA and protein levels of MMP9 after human keratinocyte cell line (HaCaT) cells were treated with 10 ng/ ml TNF-α or 2. 5 μmol/ L DAC/ 300 nmol/ L TSA. Bisulfite sequencing PCR ( BSP) and Methylation-sensitive high-resolution melt analysis ( Ms-HRM) were used to detect significantly differentially demethylated CpG sites in the human MMP9 promoter region in cells exposed to TNF-α. Different sites methylation constructs of promoter-luciferase reporter gene were made to detect the influences of site-specific DNA demethylation on transcription activity of MMP9 promoter. Results Compared with PBS-treated control, TNF-α significantly increased the expression of MMP9 in HaCaT cells for indicated culture duration ( P < 0. 05 ). Real time PCR, Western blot, and ELISA analysis demonstrated that the mRNA and protein levels of MMP9 were increased initially, followed by a decline with prolonged incubation time. After TNF-α treatment, varied degrees of DNA demethylation occurred at 10 CpG sites in the promoter of MMP9, and the changes at the -36 bp site were statistically significant (P<0. 05). The demethylation at the -36 bp site greatly increased the transcription activity of MMP9. Conclusion TNF-α promotes MMP9 expression in HaCaT cells through inducing -36 bp site DNA demethylation on the promoter of MMP9.

Objective To investigate the clinical significance of anticardiolipin antibodies (ACA)in patients with hemorrhagic fever with renal syndrome(HFRS).Methods The serum ACA- IgM and ACA-IgG in 175 cases of renal diseases and 50 healthy cases were detected by enzyme-linked immunosorbent assay(ELISA). The positive rates of ACA were calculated and compared among different groups.Results The positive rates of ACA-IgM and ACA-IgG in control group were both 0 (0/50),while they were 85.2%(46/54)and 24.1%(13/54)respectively in patients with HERS. 25.0%(13/52)and 21.2%(11/52)respectively in patients with lupus nephritis and 13.0%(9/69) and 8.7%(6/69)respectively in patients with nephrotic syndrome.The poshive rate of ACA lgM in patients with HFRS was higher than that in control group,lupus nephritis group and nephritic syn- drome group(P

[Objective]To explore the relationship between peripheral arterial disease(PAD)and diabetic retinopathy(DR)in type 2 diabetes patients.[Methods]A total of 99 patients diagnosed with PAD were classified into grade 1-3 by their total scores of peripheral arterial stenosis assessed by color doppler ultrasound examinations,where the degree of stenosis 30% ~ 49% scored 0, 50%~99%scored 1,lumen occlusion(i.e. degree of stenosis 100%)scored 2,and therefore the total score 0-2 was categorized into Grade 1 ,3~4 into Grade 2 ,5~12 into Grade 3. The bilateral anterior tibial artery ,posterior tibial artery and dorsalis pedis artery of these patients were analyzed. The presence of diabetic retinopathy(DR)was graded from retinal photographs using a standard protocol.[Results]Among 99 cases of type 2 diabetic patients with peripheral arterial disease ,58.6%of them were male with average age of 67.3 ± 7.9 years old. Patients of Grade 1,Grade2,Grade 3 lesion accounted for 45.4%,30.3%,24.2%,respectively. Age, gender,smoking history,SBP,DBP,BMI,FBG,TC,TG,LDL-C,HDL-C,HbA1C among 3 groups were not statistically signifi-cant. The associations of DM duration and HbA1C value were significantly larger in DR than in PAD. The proportion of DR patients increased with the severity degree of PAD(p for trend=0.004). Degree of stenosis Grade 2 and Grade 3 could be predictive for DR.[Conclusions]DR is associated with the severity degree of PAD in type 2 diabetes patients as evaluated by duplex ultrasonography.Degree of stenosis Grade 2 and 3 could be used for screening or finding DR. Strategies for optimum treatment and early prevention are needed.

OBJECTIVETo investigate the technological parameters of the purification process of total flavones from Smilax china with macroporous absorption resin.

METHODThe technical process for purification of total flavones with the optimum macroporous absorption resin was screened by yield of total flavones product.

RESULTThe D140 macroporous absorption resin had the best separating efficiency when the flavones content in the liquid was 0.5 g x mL(-1) equivalent to raw material, the volume of drug 18 BV (resin bad volume) with the adsorption-power 2 BV x h(-1), and the volume of 60% (mL x mL(-1)) ethanol as eluant 5-10 BV (resin bad volume) with desorption-power 1 BV x h(-1). The obtained flavones product has total flavones recovery rate of 84.72%.

CONCLUSIONThe treatment of regenerated resin is easy, this method is advisable.

Absorption ; Flavones ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Resins, Synthetic ; chemistry ; Rhizome ; chemistry ; Smilax ; chemistry ; Technology, Pharmaceutical ; methods

The effect of globular adiponectiin (gAd)on the function of NIT-1 cells under high glucose medium was investigated. The results showed that gad could completely block the increase of NADPH oxidase components p47phox expression and recover mRNA expression of pancreatic duodenal homeobox-I ,paired box gene 6,glucose transpoter 2,and glucokinase except neurogenic differentiation factor 1 (P<0.05 or P<0.01). Whereas,impaired insulin secretion and mRNA expression at high glucose concentration were not significantly improved by gAd.

Results There was no significant difference in the distribution of genotypes and allele frequencies of HSP 70-1 and HSP70-hom between case group and control group (P>0.05), but there was significant difference in the distribution of genotypes and allele frequencies of HSP70-2 between the two groups(P<0.05).There was no significant difference in the distribution of genotypes and allele frequencies of HSP70-1,HSP70-2 and HSP70-hom between Hui and Han people (P>0.05)or in the distribution of genotypes of HSP 70-2 between males and females in case group, but there was significant difference in the distribution of allele frequencies of HSP 70-2 in case group .There was no significant difference in the distribution of genotypes and allele frequencies of HSP 70-1 and HSP70-hom between males and females in case or control group .Logistic analysis confirmed that the WC, TG, TC, LDL-C, SBP, family history of diabetes and G allele were risk factors of T 2DM. Conclusion GG genotype and G allele of HSP 70-2 (+1267 ) SNP may be genetic markers for the susceptibility of type 2 diabetes.There is no significant difference in the distribution of genotypes and allele frequencies of HSP 70-1,HSP70-2 and HSP70-hom between Hui and Han nationalities .Family history of diabetes, LDL-C, TC, and G allele of HSP70-2 (+1267) SNP are the main risk factors of T2DM while HDL-C is a protective factor.

Objective To observe the clinical curative effect of drug combined with psychological treatment on patients with sleep disorders after stroke.Methods A total of 180 cases of eligible patients with sleep disorders were included and randomly di-vided into control group and treatment group.Control group were treated with conventional vascular drug(estazolam,2 mg,before sleep);treatment group increased anxiolytic and sedative hypnotic drugs(venlafaxine,dexzopiclone)compared to control group.Af-ter 14 days,sleep quality,NIHSS were compared both within the treatment group and between the two groups.Results The Bar-thel index,NIHSS and PSQI of two groups had significant differences,and those index of treatment group before and after treat-ment had significant differences too(P <0.05).Conclusion Combined conventional cerebrovascular drugs with Venlafaxine,dexzo-piclone as well as psychological treatment significantly improved sleep quality of patients with acute stroke.

OBJECTIVETo evaluate the curative effect of Qingkailing Injection (QKL) for treatment of children's respiratory syncytial virus pneumonia of phlegm-heat obstructing Fei syndrome pattern (SVP-PH) depending upon main symptom assessment.

METHODSA Chinese-Western medicine comparative trial was conducted on 206 children with SVP-PH in two groups treated with Ribavirin injection plus compound guaiacol potassium sul-fonale oral solution (as control group) and QKL injection plus Ertong Qingfei oral liquid (as treated group) respectively, for 10 days. The curative effectiveness on four main symptoms (fever, cough, sputum and short breath) were evaluated at different time-points.

RESULTSThe effectiveness in the treated group at various time-points was superior to that in the control group, showing the earlier initiating time (on the 4th day) and the preponderances on cough and sputum ran all through the whole course.

CONCLUSIONChinese medicine shows a multi-target effect in treating children's SVP-HP.

Child ; Diagnosis, Differential ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Guaiacol ; analogs & derivatives ; therapeutic use ; Humans ; Infusions, Intravenous ; Male ; Medicine, Chinese Traditional ; Phytotherapy ; Pneumonia, Viral ; drug therapy ; Respiratory Syncytial Virus Infections ; drug therapy ; Ribavirin ; therapeutic use